Reversal of Olfactory Disturbance in Allergic Rhinitis Related to OMP Suppression by Intranasal Budesonide Treatment

PURPOSE: We evaluated the severity of olfactory disturbance (OD) in the murine model of allergic rhinitis (AR) and local allergic rhinitis (LAR) in mice. We also investigated the therapeutic effect of an intranasal steroid on OD. METHODS: Forty BALB/c mice were divided into 5 groups (n = 8 for each). The control group was sensitized intraperitoneally (i.p.) and challenged intranasally (i.n.) with saline. Mice in the AR group got i.p. and i.n. ovalbumin (OVA) administration for AR induction. The LAR group was challenged i.n. with 1% OVA for inducing local nasal allergic inflammation, without inducing the systemic allergy. The OD group got an i.p. methimazole administration (75 mg/kg) to induce total destruction of olfactory mucosa. Mice in the intranasal budesonide group received i.n. budesonide (12.8 μg per time, 30 minutes after the i.n. OVA challenge) while using OVA to cause systemic allergies. We conducted a buried-food pellet test to functionally assess the degree of OD in each group by measuring the time taken until finding hidden food. We evaluated the damage to olfactory epithelium using histopathologic evaluation and compared the degree of olfactory marker protein (OMP) expression in olfactory epithelium using immunofluorescent staining. RESULTS: Mice of the AR (81.3 ± 19.8 seconds) and LAR groups (66.2 ± 12.7 seconds) spent significantly more time to detect the pellets than the control group (35.6 ± 12.2 seconds, P < 0.01). After treatment, the intranasal budesonide group exhibited significantly better results (35.8 ± 11.9 seconds) compared with the AR and LAR groups (P < 0.01). The AR and LAR groups showed considerable olfactory epithelial damage and suppression of OMP expression compared with the control group. In the intranasal budesonide group, the olfactory lesions and OMP expression had improved substantially. CONCLUSIONS: OD may be caused by olfactory epithelial damage and suppression of OMP expression in nasal allergic inflammation and could be reversed using an intranasal steroid.

Gene Expression Profile of Olfactory Transduction Signaling in an Animal Model of Human Multiple Sclerosis

Olfactory dysfunction occurs in multiple sclerosis in humans, as well as in an animal model of experimental autoimmune encephalomyelitis (EAE). The aim of this study was to analyze differentially expressed genes (DEGs) in olfactory bulb of EAE-affected mice by next generation sequencing, with a particular focus on changes in olfaction-related signals. EAE was induced in C57BL/6 mice following immunization with myelin oligodendrocyte glycoprotein and adjuvant. Inflammatory lesions were identified in the olfactory bulbs as well as in the spinal cord of immunized mice. Analysis of DEGs in the olfactory bulb of EAE-affected mice revealed that 44 genes were upregulated (and which were primarily related to inflammatory mediators), while 519 genes were downregulated; among the latter, olfactory marker protein and stomatin-like 3, which have been linked to olfactory signal transduction, were significantly downregulated (log2 [fold change] >1 and p-value < 0.05). These findings suggest that inflammation in the olfactory bulb of EAE-affected mice is associated with the downregulation of some olfactory signal transduction genes, particularly olfactory marker protein and stomatin-like 3, which may lead to olfactory dysfunction in an animal model of human multiple sclerosis.

Transplantation of Neural Stem Cells in Anosmic Mice

OBJECTIVES: Treating olfactory dysfunction is a challenge for physicians. One of the therapeutic options could be transplantation of stem cells. In this study, neural stem cells were transplanted into anosmic mice. METHODS: Neural stem cells were generated from the olfactory bulb of green fluorescent protein (GFP)-transgenic C57BL6 mice. Anosmia were induced by injection of intraperitoneal 3-methylindole. The neural stem cells were transplanted transnasally on the next day. The olfactory function was evaluated by a food-finding test once a week. The olfactory neuroepithelium was harvested for histologic examination and protein analysis at 4 weeks. RESULTS: Twenty-five percent (6/24) of the control mice that were not transplanted with neural stem cells survived at 4 weeks while 67% (8/12) of the transplanted mice survived (P=0.029). The food finding test showed that the transplanted mice resumed finding food at 3 weeks while the control mice resumed finding food at 4 weeks. GFP-positive cells were observed in the olfactory neuroepithelium of the transplanted mice. Western blotting revealed that the olfactory marker protein expression was significantly lower in the control mice than that in the transplanted mice. CONCLUSION: This study demonstrated that improvement of mouse survival was achieved and recovery of olfactory function was promoted by transnasal transplantation of neural stem cells in the anosmic mouse model. These results indicate that stem cells might be one of the future modalities for treating olfactory impairment.

Effect of Systemic Steroid on the Olfactory Epithelium Injured by 3-Methylindole in Mice

BACKGROUND AND OBJECTIVES: Various chemicals can affect the function of olfaction and steroids have been used for the treatment of olfactory dysfunction. In this study, we investigated the effect of chronic dexamethasone treatment on olfactory epithelium injured by 3-methylindole (3-MI). SUBJECTS AND METHODS: 0.75 mg/kg of dexamethasone and 0.15 mL of normal saline were administered to each of the 12 mice belonging to the experimental and control group respectively every other day from 1 week, before a single intraperitoneal administration of 175 mg/kg 3-MI, to 4 weeks after 3-MI injection. Three mice from each group were sacrificed every week, and olfactory epithelium was examined after H & E and immunohistochemical staining. RESULTS: On H & E staining, the height of the olfactory epithelium and the polarity of the cells showed no difference between the two groups. On olfactory marker protein (OMP) staining, the number of OMP-immunoreactive (IR) olfactory receptor cells was significantly increased in the experimental groups from 2 and 4 weeks after 3-MI injection compared with the control group. On proliferating cell nuclear antigen (PCNA) staining, PCNA-IR basal cells were significantly reduced in groups that received dexamethasone from 2 weeks to 3 weeks after injection compared with the control group. CONCLUSION: Dexamethasone shows no protective effect in early necrosis of olfactory epithelium by 3-MI, but showed positive effect on the regeneration of the olfactory receptor cells of olfactory epithelium.

Histopathological Changes of Olfactory Epithelium in the Rats Exposed to Cigarette Smoke / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Although exposure to cigarette smoke has been reported to be associated with olfactory dysfunction, the pathophysiology is poorly understood. The purpose of this study is to evaluate the histopathological change of olfactory epithelium according to exposure duration of cigarette smoke and to investigate the effects of cigarette smoke on the olfactory epithelium. SUBJECTS AND METHOD: Thirty-six healthy Sprague-Dawley rats were divided into 2 groups. Experimental groups (n=28) were exposed to cigarette smoke during 2.5 hours (total 5 cigarettes) per one day for 5 days, 11 days and 3, 4, 5, 6, 9 weeks. Control group (n=8) was placed at the same room without smoke exposure and sacrificed at 4 and 9 weeks. Histopathological changes of olfactory epithelium through light microscope and immunohistochemistric findings of olfactory marker protein (OMP) through confocal microscope were observed. The numbers of cells in olfactory epithelium were counted at each period. RESULTS: From 5 days of cigarette smoke exposure, defection of epithelial cells, random arrangement of olfactory receptor cells and decreased thickness of olfactory epithelium were noticed. Numbers of olfactory receptor cells were significantly decreased at 11 days and 3 weeks after smoke exposure, and this finding of decreased number of olfactory receptor cells were continued until 9 weeks of exposure. Numbers of OMP-positive olfactory receptor cells were continuously decreased according to exposure duration. CONCLUSION: The olfactory epithelial cells could be directly affected by cigarette smoke. The numbers of olfactory epithelial cells including olfactory receptor cells were continuously decreased until 9 weeks after cigarette smoke exposure.

Immunohistological Study of the Effects of Intranasal ZnSO4 Instillation on the Mouse Olfactory Epithelium and Olfactory Bulb / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The purpose of this study was to evaluate the time course of functional and anatomical recovery of the olfactory epithelium (OE) and olfactory bulb (OB) following intranasal instillation of 1% ZnSO4 in mice. MATERIALS AND METHOD: Two groups of mice, normal control group (intranasal instillation of normal saline, n=6) and experimental group (intranasal instillation of ZnSO4, n=18), were studied. Tissues of olfactory pathways were obtained at 1, 3 and 5 weeks after bilateral intranasal instillation of 1%, 100 microliter ZnSO4 solution, and processed for immunohistochemistry using antisera, olfactory marker protein (OMP), proliferating cell nuclear antigen (PCNA) and tyrosine hydroxylase (TH) to evaluate the olfactory regeneration. For histological study, OE thickness stained with hematoxylin-eosin was analyzed. RESULTS: At 1 week after ZnSO4 intranasal instillation, the lowest peak of OMP expression in OE appeared. Then the number of OMP-positive cells increased progressively at weeks. However, PCNA expression in OE showed quite the opposite. In the corresponding OB at 1 week, there was decrease of TH-positive cells and at 3 weeks, there was few TH-positive cells. At 5 weeks, there was increase in the number of TH-positive cells again. OE thickness was reduced to 20% of control OE at 1 week, and was significantly recovered to 80% of control OE at 5 weeks. CONCLUSION: Our results demonstrated that intranasal instillation of 1% ZnSO4 to mice produces a brief, reversible but essentially complete destruction of functional connection from the olfactory epithelium to the main olfactory bulb.

The Effect of Superior Cervical Ganglionectomy on Recovery of Olfaction in Induced Anosmic Mice / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: This study was undertaken to evaluate the effect of superior cervical ganglionectomy (SCG) on anosmia, which is peripherally induced in the mice. MATERIALS AND METHOD: Three groups of mice (BCF1) were studied: normal control (nasal instillation of saline, n=6); zinc sulfate group (nasal instillation of 64 mM zinc sulfate, n=25); SCG group (superior cervical ganglionectomy after nasal instillation of 64 mM zinc sulfate, n=25). Tissues of olfactory mucosa were obtained at 1, 2, 3, 4 and 7 weeks after instillation of zinc sulfate, and processed for immunohistochemistry using antisera to olfactory marker protein (OMP) to evaluate the olfactory regeneration. RESULTS: No OMP-positive cells were observed in the first two weeks after the instillation of zinc sulfate in both zinc sulfate group and the SCG group. However, the OMP-positive cells appeared first at 3 weeks after the instillation in both groups, and gradually increased in number at 4 and 7 weeks. In the SCG group, the increase of OMP-positive cells was significantly greater than those of the zinc sulfate group. The number of OMP-positive cells in the SCG group at 7 weeks was almost similar to that of the normal control group. CONCLUSION: SCG enhances regeneration of olfactory receptor cells at 3 weeks after injury. It was inferred from the above results that SCG has a significant effect on the regeneration of olfactory receptor cells and we suggest that SCG could be an effective treatment modality for olfactory dysfunction.

The Effect of Topical Steroid Nasal Instillation in Induced Anosmic Mice / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The purpose of this study was to evaluate the effect of intranasal Mometasone furoate instillation into the nasal cavity of mice which had peripherally induced anosmia. SUBJECTS AND METHOD: Three groups of mice were studied: normal control group (nasal instillation of normal saline, n=6), Mometasone furoate non-instillation group (no treatment after nasal instillation of zinc sulfate, n=12), and Mometasone furoate instillation group (daily mometasone furoate instillation after nasal instillation of zinc sulfate, n=12). Tissues of olfactory mucosa were obtained on 1, 2, 3, 4 weeks after the instillation of zinc sulfate, and processed for immunohistochemistry using antisera to olfactory marker protein (OMP) for evaluation of olfactory regeneration. RESULTS: No OMP-positive cells were observed in the first week after the instillation of zinc sulfate in both groups. However, OMP-positive cells began appearing in the second week in both groups and gradually increased as time goes by. In the Mometasone furoate instillation group, the increase of OMP-positive cells was significantly greater than that of Mometasone furoate non-instillation group. CONCLUSION: Mometasone furoate instillation enhances regeneration of olfactory receptor cells after injury. Mometasone furoate instillation can be suggested as an effective treatment modality for olfactory dysfunction.